新疆4种山楂果实中总黄酮的纯化与抗氧化活性考察

目的:优选大孔树脂纯化4种新疆山楂果实总黄酮的工艺条件,评价其抗氧化活性。方法:以总黄酮纯度为评价指标,比较13种大孔树脂对4种山楂果实总黄酮的吸附分离效果,考察上样流速、洗脱剂浓度对4种山楂总黄酮大孔树脂纯化工艺的影响。以清除2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)和二苯代苦味酰基(DPPH)自由基的方法测定样品的抗氧化活性,利用HPLC测定样品芦丁和金丝桃苷的含量,流动相乙睛-0.4%甲酸溶液(16:84),检测波长360 nm。结果:选择HPD722型大孔树脂纯化总黄酮,最佳纯化工艺条件为上样流速0.84 mL·min^-1,总黄酮经树脂充分吸附后,加40%乙醇动态洗脱。经HPD722型大孔吸附树脂纯化后,4种山楂的总黄酮、芦丁、金丝桃苷质量分数和抗氧化活性分别提高了1.9~3.3,2.4~5.1,2.6~3.7,2.0~4.6倍。结论:HPD722型大孔树脂可有效地对4种新疆山楂果实总黄酮进行富集纯化,为这4种药材资源的充分利用提供参考。

Purification and Antioxidant Activity of Total Flavonoids from Four Crataegus Fruits in Xinjiang

Objective: To optimize purification process of total flavonoids from four Crataegus fruits in Xinjiang by macroporous resin and evaluate its antioxidant activity. Method: With purity of total flavonoids as index, 13 macroporous resins for adsorption and separation of total flavonoids were compared.Purification process of total flavonoids was optimized by screening flow velocity of sample, concentration of eluent.Antioxidant activity was determined by 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid diammonium salt (ABTS) and 2, 2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method.Contents of rutin and hyperin were determined by HPLC with mobile phase of acetonitrile-0.4% formic acid (16:84) and detection wavelength at 360 nm. Result: HPD722 resin offered the best adsorption and desorption capacity for total flavonoids.Optimal purification process was as follows:flow velocity of sample 0.84 mL·min^-1, after reaching adsorption balance, dynamically eluted with 40% ethanol.After separated and purified by HPD722 resin, contents of total flavonoids, rutin, hyperin and antioxidant capacity increased 1.9-3.3, 2.4-5.1, 2.6-3.7, 2.0-4.6 times, respectively. Conclusion: Total flavonoids from four C. fruits can be effectively purified and enriched by HPD-722 macroporous resin.